EP2388329A2 - Fusion proteins the process to preparation and utilization in expression systems of recombinant proteins - Google Patents
Fusion proteins the process to preparation and utilization in expression systems of recombinant proteins Download PDFInfo
- Publication number
- EP2388329A2 EP2388329A2 EP09805852A EP09805852A EP2388329A2 EP 2388329 A2 EP2388329 A2 EP 2388329A2 EP 09805852 A EP09805852 A EP 09805852A EP 09805852 A EP09805852 A EP 09805852A EP 2388329 A2 EP2388329 A2 EP 2388329A2
- Authority
- EP
- European Patent Office
- Prior art keywords
- protein
- fragment
- proteins
- fusion
- amino acid
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Withdrawn
Links
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Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
- C12N15/62—DNA sequences coding for fusion proteins
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/43504—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from invertebrates
- C07K14/43536—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from invertebrates from worms
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/52—Cytokines; Lymphokines; Interferons
- C07K14/54—Interleukins [IL]
- C07K14/5409—IL-5
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P21/00—Preparation of peptides or proteins
- C12P21/02—Preparation of peptides or proteins having a known sequence of two or more amino acids, e.g. glutathione
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
- C07K2319/20—Fusion polypeptide containing a tag with affinity for a non-protein ligand
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
- C07K2319/35—Fusion polypeptide containing a fusion for enhanced stability/folding during expression, e.g. fusions with chaperones or thioredoxin
Definitions
- the present invention refers to fusion proteins comprising in their structure: (1) part or the full amino acid sequence of calcium binding proteins excreted/secreted by the adult worm of Fasciola hepatica; and an unrelated protein or protein fragment of interest.
- a first aspect of the present invention refers to a fusion protein comprising an amino acid sequence having the structure of the H fragment or of the calcium binding proteins excreted-secreted by the adult worm of Fasciola hepatica, followed by an amino acid sequence structurally similar to an unrelated protein fragment or protein.
- the addition of the fusion tag (H fragment or calcium binding protein) to the unrelated protein can be made at the N or C-terminal position.
- the invention relates to constructs containing the H fragments, or H fragment-containing antigens, in order to solubilize the fusion polypeptides and increase their production.
- the same methodology may be used in other protein production systems, namely fungi or eukaryotic systems.
- Protein quantification was performed by Bradford assay with the Protein Assay (Biorad) reagent diluted 1:5, and the optical density was read at a wavelength of 595 nm.
- the calibration curve was obtained from optical density readings, at 595 nm, of solutions of defined bovine sera albumin (BSA) concentrations with this reagent.
- Protein expression with the recombinant antigen was also evaluated by electrophoresis ( Figures 3 , 4 , and 5 ) under denaturating conditions (SDS-PAGE Tris-Tricine).
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Genetics & Genomics (AREA)
- Zoology (AREA)
- Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- Wood Science & Technology (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Biomedical Technology (AREA)
- General Engineering & Computer Science (AREA)
- Biophysics (AREA)
- Biotechnology (AREA)
- Gastroenterology & Hepatology (AREA)
- Microbiology (AREA)
- Toxicology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Medicinal Chemistry (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Physics & Mathematics (AREA)
- Plant Pathology (AREA)
- Peptides Or Proteins (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| PT104331A PT104331A (pt) | 2009-01-13 | 2009-01-13 | Protenas de fuso, processo para a sua preparaão e sua utilizaão em sistemas de expresso de protenas recombinantes |
| PCT/IB2009/055647 WO2010082097A2 (pt) | 2009-01-13 | 2009-12-10 | Proteínas de fusão, processo para a sua preparação e sua utilização em sistemas de expressão de proteínas recombinantes |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| EP2388329A2 true EP2388329A2 (en) | 2011-11-23 |
Family
ID=41818885
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| EP09805852A Withdrawn EP2388329A2 (en) | 2009-01-13 | 2009-12-10 | Fusion proteins the process to preparation and utilization in expression systems of recombinant proteins |
Country Status (11)
| Country | Link |
|---|---|
| US (1) | US20120165508A1 (zh) |
| EP (1) | EP2388329A2 (zh) |
| CN (1) | CN102348800B (zh) |
| AU (1) | AU2009337517A1 (zh) |
| BR (1) | BRPI0924214A2 (zh) |
| CA (1) | CA2749571A1 (zh) |
| MX (1) | MX2011007492A (zh) |
| PT (1) | PT104331A (zh) |
| SG (1) | SG174348A1 (zh) |
| WO (1) | WO2010082097A2 (zh) |
| ZA (1) | ZA201105921B (zh) |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108912221A (zh) * | 2018-07-31 | 2018-11-30 | 成都英普博集生物科技有限公司 | 用于生产重组融合蛋白的辅助蛋白、编码基因、重组融合蛋白、重组表达载体及制备方法 |
| CN112941058B (zh) * | 2021-04-02 | 2023-12-05 | 重庆科润生物医药研发有限公司 | 一种重组溶组织梭菌ii型胶原酶及其制备方法和应用 |
| CN117801124A (zh) * | 2024-02-29 | 2024-04-02 | 天津凯莱英生物科技有限公司 | 利西那肽前体的融合蛋白及其应用 |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2008002166A2 (en) * | 2006-06-27 | 2008-01-03 | Instituto Nacional De Saude Dr. Ricardo Jorge, I.P. | Diagnosis of fasciolosis by skin test (intradermoreaction) using the antigen fh8 (fasciolin). |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5270181A (en) * | 1991-02-06 | 1993-12-14 | Genetics Institute, Inc. | Peptide and protein fusions to thioredoxin and thioredoxin-like molecules |
| WO1997001627A1 (en) | 1995-06-27 | 1997-01-16 | Igen International, Inc. | High-level expression and efficient recovery of ubiquitin fusion proteins from escherichia coli |
| KR100890579B1 (ko) | 2002-08-19 | 2009-04-27 | 프로테온 주식회사 | Ｒｎａ 결합 단백질의 유전자를 융합파트너로 이용한재조합 단백질의 제조방법 |
-
2009
- 2009-01-13 PT PT104331A patent/PT104331A/pt not_active IP Right Cessation
- 2009-12-10 WO PCT/IB2009/055647 patent/WO2010082097A2/pt active Application Filing
- 2009-12-10 CN CN200980157975.2A patent/CN102348800B/zh not_active Expired - Fee Related
- 2009-12-10 AU AU2009337517A patent/AU2009337517A1/en not_active Abandoned
- 2009-12-10 CA CA2749571A patent/CA2749571A1/en not_active Abandoned
- 2009-12-10 US US13/144,493 patent/US20120165508A1/en not_active Abandoned
- 2009-12-10 MX MX2011007492A patent/MX2011007492A/es unknown
- 2009-12-10 BR BRPI0924214A patent/BRPI0924214A2/pt not_active IP Right Cessation
- 2009-12-10 SG SG2011065612A patent/SG174348A1/en unknown
- 2009-12-10 EP EP09805852A patent/EP2388329A2/en not_active Withdrawn
-
2011
- 2011-08-12 ZA ZA2011/05921A patent/ZA201105921B/en unknown
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2008002166A2 (en) * | 2006-06-27 | 2008-01-03 | Instituto Nacional De Saude Dr. Ricardo Jorge, I.P. | Diagnosis of fasciolosis by skin test (intradermoreaction) using the antigen fh8 (fasciolin). |
Non-Patent Citations (3)
| Title |
|---|
| See also references of WO2010082097A2 * |
| SILVA ELISABETE ET AL: "A recombinant antigen recognized by Fasciola hepatica-infected hosts", THE JOURNAL OF PARASITOLOGY, AMERICAN SOCIETY OF PARASITOLOGISTS, US, vol. 90, no. 4, 1 August 2004 (2004-08-01), pages 746 - 751, XP008087125, ISSN: 0022-3395, DOI: 10.1645/GE-136R * |
| SOFIA J. COSTA ET AL: "The novel Fh8 and H fusion partners for soluble protein expression in Escherichia coli: a comparison with the traditional gene fusion technology", APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, vol. 97, no. 15, 20 November 2012 (2012-11-20), pages 6779 - 6791, XP055091272, ISSN: 0175-7598, DOI: 10.1007/s00253-012-4559-1 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CA2749571A1 (en) | 2010-07-22 |
| BRPI0924214A2 (pt) | 2015-09-15 |
| ZA201105921B (en) | 2012-10-31 |
| US20120165508A1 (en) | 2012-06-28 |
| CN102348800A (zh) | 2012-02-08 |
| MX2011007492A (es) | 2011-11-18 |
| CN102348800B (zh) | 2015-07-22 |
| SG174348A1 (en) | 2011-10-28 |
| WO2010082097A2 (pt) | 2010-07-22 |
| PT104331A (pt) | 2010-07-13 |
| WO2010082097A4 (pt) | 2011-03-17 |
| WO2010082097A3 (pt) | 2010-12-23 |
| AU2009337517A1 (en) | 2011-09-01 |
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